LOCUS       sequence-254281-       10867 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AMA1 plasmid with argB selection marker, and USER cassette 
            (PacI/Nt.BbvCI).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10867)
  AUTHORS   Vanegas KG, Jarczynska ZD, Strucko T, Mortensen UH
  TITLE     Cpf1 enables fast and efficient genome editing in Aspergilli.
  JOURNAL   Fungal Biol Biotechnol. 2019 May 1;6:6. doi: 
            10.1186/s40694-019-0069-6. eCollection 2019.
  PUBMED    31061713
REFERENCE   2  (bases 1 to 10867)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Fungal Biol
            Biotechnol."; date: "2019-05-1"; volume: "6:6. doi"; pages: " 
            10.1186/s40694-019-0069-6. eCollection 2019"
COMMENT     Sequence Label: pAC573
FEATURES             Location/Qualifiers
     source          1..10867
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     622..641
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(860..951)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     993..1012
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1018..1037
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     protein_bind    1244..1277
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     rep_origin      1590..6838
                     /label=AMA1
                     /note="replication origin from Aspergillus nidulans 
                     (Aleksenko and Clutterbuck, 1996)"
     protein_bind    complement(9367..9400)
                     /label=FRT (minimal)
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="supports FLP-mediated excision but not integration 
                     (Turan and Bode, 2011)"
     primer_bind     complement(9870..9886)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(9985..10002)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(10156..10744)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(10236..10255)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(10866..10867,1..790,791..859))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag
       61 ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca
      121 gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca gcaataaacc
      181 agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt
      241 ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg
      301 ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca
      361 gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg
      421 ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca
      481 tggttatggc agcgctacat aattctctta ctgtcatgcc atccgtaaga tgcttttctg
      541 tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct
      601 cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca
      661 tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca
      721 gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg
      781 tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac
      841 ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt tatcagggtt
      901 attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggtca
      961 gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata cctgaatatg
     1021 gctcataaca ccccttgttt gcctggcggc agtagcgcgg tggtcccacc tgaccccatg
     1081 ccgaactcag aagtgaaacg ccgtagcgcc gatggtagtg tggggactcc ccatgcgaga
     1141 gtagggaact gccaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     1201 cccgggctaa ttatggggtg tcgcccttat tcgactctat agtgaagttc ctattctcta
     1261 gaaagtatag gaacttctga agtggggatt taaatgcggc cgtgctgagg gtttaatcga
     1321 cgaagcagct gacggccagt gccaagctta acgcgtaccg ggcccagtat atgttccgca
     1381 gatgactgga gctctgccat acgtgccctc tcaagcacca tttgttccat ctacagagac
     1441 tagtcaccaa ctagtctatc aagactcaca tggtacattg ctgagaccaa ctgaccagag
     1501 gcagggtagc ggattgacgg ctccatctcc ttcacttaca aggtctattg aaagcccttt
     1561 agcatcacca agcggagaat agattgttaa gcttattttt tgtatactgt tttgtgatag
     1621 cacgaagttt ttccacggta tcttgttaaa aatatatatt tgtggcgggc ttacctacat
     1681 caaattaata agagactaat tataaactaa acacacaagc aagctacttt agggtaaaag
     1741 tttataaatg cttttgacgt ataaacgttg cttgtattta ttattacaat taaaggtgga
     1801 tagaaaacct agagactagt tagaaactaa tctcaggttt gcgttaaact aaatcagagc
     1861 ccgagaggtt aacagaacct agaaggggac tagatatccg ggtagggaaa caaaaaaaaa
     1921 aaacaagaca gccacatatt agggagacta gttagaagct agttccagga ctaggaaaat
     1981 aaaagacaat gataccacag tctagttgac aactagatag attctagatt gaggccaaag
     2041 tctctgagat ccaggttagt tgcaactaat actagttagt atctagtctc ctataactct
     2101 gaagctagaa taacttacta ctattatcct caccactgtt cagctgcgca aacggagtga
     2161 ttgcaaggtg ttcagagact agttattgac tagtcagtga ctagcaataa ctaacaaggt
     2221 attaacctac catgtctgcc atcaccctgc acttcctcgg gctcagcagc cttttcctcc
     2281 tcattttcat gctcattttc cttgtttaag actgtgacta gtcaaagact agtccagaac
     2341 cacaaaggag aaatgtctta ccactttctt cattgcttgt ctcttttgca ttatccatgt
     2401 ctgcaactag ttagagtcta gttagtgact agtccgacga ggacttgctt gtctccggat
     2461 tgttggagga actctccagg gcctcaagat ccacaacaga gccttctaga agactggtca
     2521 ataactagtt ggtctttgtc tgagtctgac ttacgaggtt gcatactcgc tccctttgcc
     2581 tcgtcaatcg atgagaaaaa gcgccaaaac tcgcaatatg gctttgaacc acacggtgct
     2641 gagactagtt agaatctagt cccaaactag cttggatagc ttacctttgc cctttgcgtt
     2701 gcgacaggtc ttgcagggta tggttccttt ctcaccagct gatttagctg ccttgctacc
     2761 ctcacggcgg atctgcataa agagtggcta gaggttataa attagcactg atcctaggta
     2821 cggggctgaa tgtaacttgc cyttcctttc tcatcgcgcg gcaagacagg cttgctcaaa
     2881 ttcctaccag tcacaggggt atgcacggcg tacggaccac ttgaactagt cacagattag
     2941 ttagcaacta gtctgcattg aatggctgta cttacgggcc ctcgccattg tcctgatcat
     3001 ttccagcttc accctcgttg ctgcaaagta gttagtgact agtcaaggac tagttgaaat
     3061 gggagaagaa actcacgaat tctcgacacc cttagtattg tggtccttgg acttggtgct
     3121 gctatatatt agctaataca ctagttagac tcacagaaac ttacgcagct cgcttgckct
     3181 tcttggtagg agtcggggtt gggagaacag tgccttcaaa caagccttca taccatgctr
     3241 cttgactagt cagggactag tcaccaagta atctagatag gacttgcctt tggcctccat
     3301 cagttccttc atagtgggag gtccattgtg caatgtaaac tccatgccgt gggagttctt
     3361 gtccttcaag tgcttgacca atatgtttct gttggcagag ggaacctgtc aactagttaa
     3421 taactagtca gaaactagta tagcagtaga ctcactgtac gcttgaggca tcccttcact
     3481 cggcagtaga cttcatatgg atggatatca ggcacgccat tgtcgtcctg tggactagtc
     3541 agtaactagg cttaaagcta gtcgggtcgg cttactatct tgaaatccgg cagcgtaagc
     3601 tccccgtcct taactgcctc gagatagtga cagtactctg gggactttcg gagatcgtta
     3661 tcgcgaatgc tcggcatact aatcgttgac tagtcttgga ctagtcccga gcaaaaagga
     3721 ttggaggagg aggaggaagg tgagagtgag acaaagagcg aaataagagc ttcaaaggct
     3781 atctctaagc agtatgaagg ttaagtatct agttcttgac tagatttaaa agagatttcg
     3841 actagttatg tacctggagt ttggatatag gaatgtgttg tggtaacgaa atgtaagggg
     3901 gaggaaagaa aaagtcggtc aagaggtaac tctaagtcgg ccattccttt ttgggaggcg
     3961 ctaaccataa acggcatggt cgacttagag ttagctcagg gaatttaggg agttatctgc
     4021 gaccaccgag gaacggcgga atgccaaaga atcccgatgg agctctagct ggcggttgac
     4081 aaccccacct tttggcgttt ctgcggcgtt gcaggcggga ctggatactt cgtagaacca
     4141 gaaaggcaag gcagaacgcg ctcagcaaga gtgttggaag tgatagcatg atgtgccttg
     4201 ttaactaggt caaaatctgc agtatgcttg atgttatcca aagtgtgaga gaggaaggtc
     4261 caaacataca cgatttggag agggcctagg tataagagtt tttgagtaga acgcatgtga
     4321 gcccagccat ctcgaggaga ttaaacacgg gccggcattt gatggctatg ttagtacccc
     4381 aatggaaacg gtgagagtcc agtggtcgca gataactccc taaattccct gagctaactc
     4441 taagtcgacc atgccgttta tggttagcgc ctcccaaaaa ggaatggccg acttagagtt
     4501 acctcttgac cgactttttc tttcctcccc cttacatttc gttaccacaa cacattccta
     4561 tatccaaact ccaggtacat aactagtcga aatctctttt aaatctagtc aagaactaga
     4621 tacttaacct tcatactgct tagagatagc ctttgaagct cttatttcgc tctttgtctc
     4681 actctcacct tcctcctcct cctccaatcc tttttgctcg ggactagtcc aagactagtc
     4741 aacgattagt atgccgagca ttcgcgataa cgatctccga aagtccccag agtactgtca
     4801 ctatctcgag gcagttaagg acggggagct tacgctgccg gatttcaaga tagtaagccg
     4861 acccgactag ctttaagcct agttactgac tagtccacag gacgacaatg gcgtgcctga
     4921 tatccatcca tatgaagtct actgccgagt gaagggatgc ctcaagcgta cagtgagtct
     4981 actgctatac tagtttctga ctagttatta actagttgac aggttccctc tgccaacaga
     5041 aacatattgg tcaagcactt gaaggacaag aactcccacg gcatggagtt tacattgcac
     5101 aatggacctc ccactatgaa ggaactgatg gaggccaaag gcaagtccta tctagattac
     5161 ttggtgacta gtccctgact agtcaagyag catggtatga aggcttgttt gaaggcactg
     5221 ttctcccaac cccgactcct accaagaagm gcaagcgagc tgcgtaagtt tctgtgagtc
     5281 taactagtgt attagctaat atatagcagc accaagtcca aggaccacaa tactaagggt
     5341 gtcgagaatt cgtgagtttc ttctcccatt tcaactagtc cttgactagt cactaactac
     5401 tttgcagcaa cgagggtgaa gctggaaatg atcaggacaa tggcgagggc ccgtaagtac
     5461 agccattcaa tgcagactag ttgctaacta atctgtgact agttcaagtg gtccgtacgc
     5521 cgtgcatacc cctgtgactg gtaggaattt gagcaagcct gtcttgccgc gcgatgagaa
     5581 aggaarggca agttacattc agccccgtac ctaggatcag tgctaattta taacctctag
     5641 ccactcttta tgcagatccg ccgtgagggt agcaaggcag ctaaatcagc tggtgagaaa
     5701 ggaaccatac cctgcaagac ctgtcgcaac gcaaagggca aaggtaagct atccaagcta
     5761 gtttgggact agattctaac tagtctcagc accgtgtggt tcaaagccat attgcgagtt
     5821 ttggcgcttt ttctcatcga ttgacgaggc aaagggagcg agtatgcaac ctcgtaagtc
     5881 agactcagac aaagaccaac tagttattga ccagtcttct agaaggctct gttgtggatc
     5941 ttgaggccct ggagagttcc tccaacaatc cggagacaag caagtcctcg tcggactagt
     6001 cactaactag actctaacta gttgcagaca tggataatgc aaaagagaca agcaatgaag
     6061 aaagtggtaa gacatttctc ctttgtggtt ctggactagt ctttgactag tcacagtctt
     6121 aaacaaggaa aatgagcatg aaaatgagga ggaaaaggct gctgagcccg aggaagtgca
     6181 gggtgatggc agacatggta ggttaatacc ttgttagtta ttgctagtca ctgactagtc
     6241 aataactagt ctctgaacac cttgcaatca ctccgtttgc gcagctgaac agtggtgagg
     6301 ataatagtag taagttattc tagcttcaga gttataggag actagatact aactagtatt
     6361 agttgcaact aacctggatc tcagagactt tggcctcaat ctagaatcta tctagttgtc
     6421 aactagactg tggtatcatt gtcttttatt ttcctagtcc tggaactagc ttctaactag
     6481 tctccctaat atgtggctgt cttgtttttt ttttttgttt ccctacccgg atatctagtc
     6541 cccttatagg ttctgttaac ctctcgggct ctgatttagt ttaacgcaaa cctgagatta
     6601 gtttctaact agtctctagg ttttctatcc acctttaatt gtaataataa atacaagcaa
     6661 cgtttatacg tcaaaagcat ttataagctt ttaccctaaa gtagcttgct tgtgtgttta
     6721 gtttataatt agtctcttat taatttgatg taggtaagcc cgccacaaat atatattttt
     6781 aacaagatac cgtggaaaaa cttcgtgcta tcacaaaaca gtatacaaaa aataagctta
     6841 acaatctatt ctccgcttgg tgatgctaaa gggctttcaa tagaccttgt aagtgaagga
     6901 gatggagccg tcaatccgct accctgcctc tggtcagttg gtctcagcaa tgtaccctgt
     6961 gagtcttgat agactagttg gtgactagtc tctgtagatg gaacaaatgg tgcttgagag
     7021 ggcacgtatg gcagagctcc agtcatctgc ggaacatata ctgggcccgg gaagatctca
     7081 tggtcatagc tgtttccgct gagggtttaa ttaagacctc agcactaggt aatatcgcgt
     7141 gcattccgcg gttttttggg gtagtcatct aatgaaacag acccggacgc agcagaggaa
     7201 gcccgcgatg actctatacc accgtacgcc gatatatcat catcgcggcg atggagaagt
     7261 ggggttgact ccgaagacac ttcaaaggag cgacgctgtt gatttgtaga cgacgcttga
     7321 tagggagaag cattattgtc gtgatgctcg cccaacagag gccgactcgc ctcatccgtc
     7381 ataacgaacg ctgtgtaaag cggagtgggg gggaaagtgt ggattgtgga gagtatgcga
     7441 tagtgttgag gctgatcaga cggcgaatcg ggccagatat gaccagttta gaggcctcat
     7501 ttgactataa tttacataaa ttagataaat agagatgaac gcatgcaata attgcagcaa
     7561 atattgatga agcgagaggt aggacgatga aggactgtga gcagttcaag gtatcagcag
     7621 agtcaagggc ctgatgcaat ggcggtgatc cgtgatcagc gaacggaagg ggcgctaact
     7681 ctgtttcttt accaatgatc ggaagctcct gctggcggac ttatgagtca ttcacgaatc
     7741 atttctcagt tatttgtgga tgccctcgtt ctgtccacaa tttctttccg ccccaagtct
     7801 tttaagttct ttaacatcta tattcttgca cttccaatgg catcccttcg ctccgtactc
     7861 aagagccaga gcttgcgaca caccgtgcga tcctactcct cgcaaaccat gcctcccgcc
     7921 tcaccctttg ctccccgcca cttcctctcc attgcggacc tctcgccctc cgagttcgca
     7981 acccttgttc gcaatgcctc ctcacacaaa cgggctatca agtcggggtc aatgccccaa
     8041 aacttgcagg gatcactcct tgggaaaact gtggccatga tcttcagcaa acgaagcacg
     8101 aggacaaggg tatctacaga aggggccgtt gtgcagatgg gaggtcatcc gatgttcttg
     8161 ggcaaggatg atatccaact aggtgtcaac gagtccctat acgacacctc cgttgtcatt
     8221 tcgtccatgg tatcctgcat tgtagcccgt gtcggtaaac atgcagaggt cgcagatctg
     8281 gcgaagcact cttcggttcc agtcatcaat gctttgtgtg actctttcca ccctctccaa
     8341 gccgtggccg atttccagac catctatgaa gcattcaccc ccaaggcgca ccacctttca
     8401 agtctagggt tggaaggatt gaagatcgct tgggtgggtg acgccaacaa cgtcctgttc
     8461 gatatggcca ttgctgctac aaaaatgggt gtcgacattg ctgtcgctac tcccaagggg
     8521 tacgaaatcc ctcctcacat gctggagctc atcaagtctg ctggagaggg tgtctcgaaa
     8581 ccaggaaagc ttctgcaaac caatattccc gaagaagcgg tcaaggacgc cgatattctg
     8641 gtcacagaca cctgggtctc tatgggccaa gaggaagaga aggctcagag gctgaaggag
     8701 tttgatggtt tccaaatcac tgctgaactc gccaagcgag gaggagctaa ggagggctgg
     8761 aagttcatgc actgtctccc gcgacaccct gaggaggtca gcgacgaggt tttctacagc
     8821 aaccggtcac ttgtcttccc tgaggctgag aaccggttat gggctgcgat ttccgccttg
     8881 gagggtttcg ttgtcaataa gggaaaaatt gaataaatat aaccaggctt ccatttaaat
     8941 atatagaggc tggcgttatc aaactgatga gttgacgggt atgagatcat tccgtcccta
     9001 aatatattta ctccgatcac gtaaaagcct gttagtagaa gcattttccc aattatcctg
     9061 accaattctt tctagcatat atcaaataac taattgacat gttctctcgc ttccttatat
     9121 tactcagagt attggaaatg gggcaaatcg cacccggtga ctttcacatg tcacgaatgc
     9181 ggagtcgtcc tagccaaggt agatccaggc ctaacacacc ccaaccctcg actctcctca
     9241 ttccatactc ttgacctcta tccagcacat tcttctgagg tttcgcaatg gctgtaggtg
     9301 gcgctgaggc atccccacta ccgcattaag acctcagcgc ggccgcaaat ttaaataaaa
     9361 tgaagtgaag ttcctatact ttctagagaa taggaacttc tatagtgagt cgaataaggg
     9421 cgacacaaaa tttattctaa atgcataata aatactgata acatcttata gtttgtatta
     9481 tattttgtat tatcgttgac atgtataatt ttgatatcaa aaactgattt tccctttatt
     9541 attttcgaga tttattttct taattctctt taacaaacta gaaatattgt atatacaaaa
     9601 aatcataaat aatagatgaa tagtttaatt ataggtgttc atcaatcgaa aaagcaacgt
     9661 atcttattta aagtgcgttg cttttttctc atttataagg ttaaataatt ctcatatatc
     9721 aagcaaagtg acaggcgccc ttaaatattc tgacaaatgc tctttcccta aactcccccc
     9781 ataaaaaaac ccgccgaagc gggtttttac gttatttgcg gattaacgat tactcgttat
     9841 cagaaccgcc cagggggccc gagcttaaga ctggccgtcg ttttacaaca cagaaagagt
     9901 ttgtagaaac gcaaaaaggc catccgtcag gggccttctg cttagtttga tgcctggcag
     9961 ttccctactc tcgccttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct
    10021 gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcaggtga
    10081 taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc
    10141 cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg
    10201 ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg
    10261 aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt
    10321 tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt
    10381 gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc cctaccgctg
    10441 cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact
    10501 ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt
    10561 cttgaagtgg tgggctaact acggctacac tagaagaaca gtatttggta tctgcgctct
    10621 gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac
    10681 cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc
    10741 tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg acgcgcgcgt
    10801 aactcacgtt aagggatttt ggtcatgagc ttgcgccgtc ccgtcaagtc agcgtaatgc
    10861 tctgctt
//
